Source partitioning using N2O isotopomers and soil WFPS to establish dominant N2O production pathways from different pasture sward compositions.

Nitrous oxide (N2O) is a potent greenhouse gas (GHG) emitted from agricultural soils and is influenced by nitrogen (N) fertiliser management and weather and soil conditions. Source partitioning N2O emissions related to management practices and soil conditions could suggest effective mitigation strategies. Multispecies swards can maintain herbage yields at reduced N fertiliser rates compared to grass monocultures and may reduce N losses to the wider environment. A restricted-simplex centroid experiment was used to measure daily N2O fluxes and associated isotopomers from eight experimental plots (7.8 m2) post a urea-N fertiliser application (40 kg N ha-1). Experimental pastures consisted of differing proportions of grass, legume and forage herb represented by perennial ryegrass (Lolium perenne), white clover (Trifolium repens) and ribwort plantain (Plantago lanceolata), respectively. N2O isotopomers were measured using a cavity ring down spectroscopy (CRDS) instrument adapted with a small sample isotope module (SSIM) for the analysis of gas samples ≤20 mL. Site preference (SP = δ15Nα - δ15Nß) and δ15Nbulk ((δ15Nα + δ15Nß) / 2) values were used to attribute N2O production to nitrification, denitrification or a mixture of both nitrification and denitrification over a range of soil WFPS (%). Daily N2O fluxes ranged from 8.26 to 86.86 g N2O-N ha-1 d-1. Overall, 34.2% of daily N2O fluxes were attributed to nitrification, 29.0% to denitrification and 36.8% to a mixture of both. A significant diversity effect of white clover and ribwort plantain on predicted SP and δ15Nbulk indicated that the inclusion of ribwort plantain may decrease N2O emission through biological nitrification inhibition under drier soil conditions (31%-75% WFPS). Likewise, a sharp decline in predicted SP indicates that increased white clover content could increase N2O emissions associated with denitrification under elevated soil moisture conditions (43%-77% WFPS). Biological nitrification inhibition from ribwort plantain inclusion in grassland swards and management of N fertiliser source and application timing to match soil moisture conditions could be useful N2O mitigation strategies.

Development and verification of a novel isotopic N2 O measurement technique for discrete static chamber samples using cavity ring-down spectroscopy.

RATIONALE: N2 O isotopomers are a useful tool to study soil N cycling processes. The reliability of such measurements requires a consistent set of international N2 O isotope reference materials to improve inter-laboratory and inter-instrument comparability and avoid reporting inaccurate results. All these are the more important given the role of N2 O in anthropogenic climate change and the pressing need to develop our understanding of soil N cycling and N2 O emission to mitigate such emissions. Cavity ring-down spectroscopy (CRDS) could potentially overcome resource requirements and technical challenges, making N2 O isotopomer measurements more feasible and less expensive than previous approaches (e.g., gas chromatography [GC] and isotope ratio mass spectrometry [IRMS]). METHODS: A combined laser spectrometer and small sample isotope module (CRDS & SSIM) method enabled N2 O concentration, δ15 Nbulk , δ15 Nα , δ15 Nß and site preference (SP) measurements of sample volumes <20 mL, such as static chamber samples. Sample dilution and isotopic mixing as well as N2 O concentration dependence were corrected numerically. A two-point calibration procedure normalised δ values to the international isotope-ratio scales. The CRDS & SSIM repeatability was determined using a reference gas (Ref Gas). CRDS & SSIM concentration measurements were compared with those obtained by GC, and the isotope ratio measurements from two different mass spectrometers were compared. RESULTS: The repeatability (mean ± 1σ; n = 10) of the CRDS & SSIM measurements of the Ref Gas was 710.64 ppb (± 8.64), 2.82‰ (± 0.91), 5.41‰ (± 2.00), 0.23‰ (± 0.22) and 5.18‰ (± 2.18) for N2 O concentration, δ15 Nbulk , δ15 Nα , δ15 Nß and SP, respectively. The CRDS & SSIM concentration measurements were strongly correlated with GC (r = 0.99), and they were more precise than those obtained using GC except when the N2 O concentrations exceeded the specified operating range. Normalising CRDS & SSIM δ values to the international isotope-ratio scales using isotopic N2 O standards (AK1 and Mix1) produced accurate results when the samples were bracketed within the range of the δ values of the standards. The CRDS & SSIM δ15 Nbulk and SP precision was approximately one order of magnitude less than the typical IRMS precision. CONCLUSIONS: CRDS & SSIM is a promising approach that enables N2 O concentrations and isotope ratios to be measured by CRDS for samples <20 mL. The CRDS & SSIM repeatability makes this approach suitable for N2 O "isotopomer mapping" to distinguish dominant source pathways, such as nitrification and denitrification, and requires less extensive lab resources than the traditionally used GC/IRMS. Current study limitations highlighted potential improvements for future users of this approach to consider, such as automation and physical removal of interfering trace gases before sample analysis.

Sward composition and soil moisture conditions affect nitrous oxide emissions and soil nitrogen dynamics following urea-nitrogen application.

Increased emissions of N2O, a potent greenhouse gas (GHG), from agricultural soils is a major concern for the sustainability of grassland agriculture. Emissions of N2O are closely associated with the rates and forms of N fertilisers applied as well as prevailing weather and soil conditions. Evidence suggests that multispecies swards require less fertiliser N input, and may cycle N differently, thus reducing N loss to the environment. This study used a restricted simplex-centroid experimental design to investigate N2O emissions and soil N cycling following application of urea-N (40 kg N ha-1) to eight experimental swards (7.8 m2) with differing proportions of three plant functional groups (grass, legume, herb) represented by perennial ryegrass (PRG, Lolium perenne), white clover (WC, Trifolium repens) and ribwort plantain (PLAN, Plantago lanceolata), respectively. Swards were maintained under two contrasting soil moisture conditions to examine the balance between nitrification and denitrification. Two N2O peaks coincided with fertiliser application and heavy rainfall events; 13.4 and 17.7 g N2O-N ha-1 day-1 (ambient soil moisture) and 39.8 and 86.9 g N2O-N ha-1 day-1 (wet soil moisture). Overall, cumulative N2O emissions post-fertiliser application were higher under wet soil conditions. Increasing legume (WC) proportions from 0% to 60% in multispecies swards resulted in model predicted N2O emissions increasing from 22.3 to 96.2 g N2O-N ha-1 (ambient soil conditions) and from 59.0 to 219.3 g N2O-N ha-1 (wet soil conditions), after a uniform N application rate. Soil N dynamics support denitrification as the dominant source of N2O especially under wet soil conditions. Significant interactions of PRG or WC with PLAN on soil mineral N concentrations indicated that multispecies swards containing PLAN potentially inhibit nitrification and could be a useful mitigation strategy for N loss to the environment from grassland agriculture.

Antimicrobial susceptibility of methicillin-resistant Staphylococcus pseudintermedius isolated from veterinary clinical cases in the UK.

Staphylococcus pseudintermedius is a leading aetiologic agent of pyoderma and other body tissue infections in dogs and cats. In recent years, an increased prevalence of methicillin-resistant S. pseudintermedius (MRSP) has been reported. Isolation of MRSP in serious infections poses a major therapeutic challenge as strains are often resistant to all forms of systemic antibiotic used to treat S. pseudintermedius -related infections. This study investigates the occurrence of MRSP from a total of 7183 clinical samples submitted to the authors' laboratories over a 15-month period. Identification was based on standard microbiological identification methods, and by S. pseudintermedius-specific nuc polymerase chain reaction (PCR). Methicillin resistance was confirmed by PBP2a latex agglutination and mecA PCR. Susceptibility against non-beta-lactam antibiotics was carried out using a disc-diffusion method according to Clinical and Laboratory Standards Institute (CLSI) guidelines. In addition, susceptibility to pradofloxacin--a new veterinary fluoroquinolone--was also investigated. SCCmec types were determined by multiplex PCR. Staphylococcus pseudintermedius was isolated from 391 (5%) samples and 20 were confirmed as MRSP from cases of pyoderma, otitis, wound infections, urinary tract infection and mastitis in dogs only. All 20 isolates were resistant to clindamycin and sulphamethoxazole/trimethoprim. Nineteen were resistant to chloramphenicol, enrofloxacin, gentamicin, marbofloxacin and pradofloxacin; additionally, seven isolates were resistant to tetracycline. Fifteen isolates carried SCCmec type II-III, four isolates had type V and one harboured type IV. To date, only a few scientific papers on clinical MRSP strains isolated from the UK have been published, thus the results from this study would provide additional baseline data for further investigations.

Quantification of phosphorus transport from a karstic agricultural watershed to emerging spring water.

The degree to which waters in a given watershed will be affected by nutrient export can be defined as that watershed's nutrient vulnerability. This study applied concepts of specific phosphorus (P) vulnerability to develop intrinsic groundwater vulnerability risk assessments in a 32 km(2) karst watershed (spring zone of contribution) in a relatively intensive agricultural landscape. To explain why emergent spring water was below an ecological impairment threshold, concepts of P attenuation potential were investigated along the nutrient transfer continuum based on soil P buffering, depth to bedrock, and retention within the aquifer. Surface karst features, such as enclosed depressions, were reclassified based on P attenuation potential in soil at the base. New techniques of high temporal resolution monitoring of P loads in the emergent spring made it possible to estimate P transfer pathways and retention within the aquifer and indicated small-medium fissure flows to be the dominant pathway, delivering 52-90% of P loads during storm events. Annual total P delivery to the main emerging spring was 92.7 and 138.4 kg total P (and 52.4 and 91.3 kg as total reactive P) for two monitored years, respectively. A revised groundwater vulnerability assessment was used to produce a specific P vulnerability map that used the soil and hydrogeological P buffering potential of the watershed as key assumptions in moderating P export to the emergent spring. Using this map and soil P data, the definition of critical source areas in karst landscapes was demonstrated.

Performance of a woodchip filter to treat dairy soiled water.

The milking process on dairy farms produces wastewaters, known as dairy soiled waters (DSW), which contain variable concentrations of nutrients. The most common method of disposal is by application to land. However, this practise can result in the pollution of nearby receiving water bodies. A laboratory study, comprising two sets of 0.5 m, 1 m and 1.5 m-deep filters loaded at two loading rates, examined the performance of woodchip filters in treating DSW. The filters comprised de-barked Sitka spruce (Picea sitchensis) woodchips. Dried DSW was reconstituted to 1% and 3% suspended solids (SS) concentrations and was applied at 28 L/m(2).d to the surface of the filters at loading rates of 280 g SS/m(2).d and 840 g SS/m(2).d, respectively. Filters were loaded for a maximum of 277 days. The filters achieved substantial decreases in SS (>99%), chemical oxygen demand (COD) (>97%) and total nitrogen (TN) (>89%). The dominant treatment mechanism appears to be physical filtration, but sorption and biological uptake likely also play a role. As the filters are aerobic, mineralisation and nitrification occur, but gaseous N losses are probably not significant. Woodchip shows potential as a filter medium for treating DSW, significantly decreasing the concentrations of SS, COD and TN.

On-farm treatment of dairy soiled water using aerobic woodchip filters.

Dairy soiled water (DSW) is produced on dairy farms through the washing-down of milking parlours and holding areas, and is generally applied to land. However, there is a risk of nutrient loss to surface and ground waters from land application. The aim of this study was to use aerobic woodchip filters to remove organic matter, suspended solids (SS) and nutrients from DSW. This novel treatment method would allow the re-use of the final effluent from the woodchip filters to wash down yards, thereby reducing water usage and environmental risks associated with land spreading. Three replicate 100 m(2) farm-scale woodchip filters, each 1 m deep, were constructed and operated to treat DSW from 300 cows over an 11-month study duration. The filters were loaded at a hydraulic loading rate of 30 L m(-2) d(-1), applied in four doses through a network of pipes on the filter surface. Average influent concentrations of chemical oxygen demand (COD), SS and total nitrogen (TN) of 5750 ± 1441 mg L(-1), 602 ± 303 mg L(-1) and 357 ± 100 mg L(-1), respectively, were reduced by 66, 86 and 57% in the filters. Effluent nutrient concentrations remained relatively stable over the study period, indicating the effectiveness of the filter despite increasing and/or fluctuating influent concentrations. Woodchip filters are a low cost, minimal maintenance treatment system, using a renewable resource that can be easily integrated into existing farm infrastructure.

Carriage of methicillin-resistant Staphylococcus pseudintermedius in small animal veterinarians: indirect evidence of zoonotic transmission.

Methicillin-resistant Staphylococcus pseudintermedius (MRSP) is increasingly reported in small animals and cases of human infections have already been described despite its recent emergence in veterinary practice. We investigated the prevalence of MRSP and methicillin-resistant Staphylococcus aureus (MRSA) among small animal dermatologists attending a national veterinary conference in Italy. Nasal swabs were obtained from 128 veterinarians, seven of which harboured MRSP (n = 5; 3.9%) or MRSA (n = 2; 1.6%). A follow-up study of two carriers revealed that MRSP persisted for at least 1 month in the nasal cavity. Methicillin-susceptible S. aureus (MSSA) was isolated from 32 (25%) conference participants, whereas methicillin-susceptible S. pseudintermedius (MSSP) was not detected, suggesting that MRSP may have a particular ability to colonize humans compared to MSSP. All isolates were characterized by spa typing. Methicillin-resistant isolates were further typed by antimicrobial susceptibility testing, SCCmec and multi-locus sequence typing. Two lineages previously associated with pets were identified among the five MRSP isolates; the European epidemic clone ST71-SCCmec II-III and ST106-SCCmec IV. One of the two MRSA isolates displayed a genotype (ST22- SCCmecIV) frequently reported in dogs and cats. MRSP isolates were resistant to more antimicrobial agents compared with MRSA isolates and displayed the typical multidrug resistance patterns of MRSP in pets. The 32 MSSA isolates belonged to 20 spa types and the most frequent types (t12, t15 and t166) were associated with common S. aureus lineages in humans (CC30 and CC45). Although low, the 3.9% MRSP carriage rate found among small animal dermatologists was surprising in consideration of the rare occurrence of S. pseudintermedius in humans, the lack of MSSP detection and the recent appearance of MRSP in Europe. As cases of human MRSP infection have been linked with pets, veterinarians should be aware of this zoonotic risk and proper preventative measures should be taken to avoid MRSP transmission from animal patients.